Interrogating Satellite Cell and Myofiber Defects and Repair in Human DMD using Single Nuclei/Single Cell RNA Sequencing of Muscle Resident Cells
Grant Award Details
Grant Type:
Grant Number:
DISC0-15921
Investigator(s):
Human Stem Cell Use:
Award Value:
$1,578,000
Status:
Active
Grant Application Details
Application Title:
Interrogating Satellite Cell and Myofiber Defects and Repair in Human DMD using Single Nuclei/Single Cell RNA Sequencing of Muscle Resident Cells
Public Abstract:
Research Objective
We will describe, for the first time, human muscle satellite cell, myofiber and immune cell dynamics due to dystrophin deficiency and AAV gene therapy in human muscle at single nuclei resolution.
Impact
These studies will elucidate satellite stem cell and myofiber defects in Duchenne and Becker Muscular Dystrophy and determine efficacy, mechanism and toxicities of exon skipping and AAV-gene therapy.
Major Proposed Activities
We will describe, for the first time, human muscle satellite cell, myofiber and immune cell dynamics due to dystrophin deficiency and AAV gene therapy in human muscle at single nuclei resolution.
Impact
These studies will elucidate satellite stem cell and myofiber defects in Duchenne and Becker Muscular Dystrophy and determine efficacy, mechanism and toxicities of exon skipping and AAV-gene therapy.
Major Proposed Activities
- Continue bioinformatic analysis of existing DMD snseq/scRNAseq dataset.
- Perform biopsies and prepare and bank new muscle tissue, PBMC and expanded satellite and T cell populations in culture and determine dystrophin expression.
- Extract nuclei for snRNAeq and cluster analyze newly acquired BMD/DMD biopsy snRNAseq in the context of our large healthy and DMD snRNAseq data reference set.
- Perform pilot experiments involving TNC/EGFR competitor interaction to follow up snRNAseq data identifying increases in TNC expression in DMD versus healthy muscle.
- Collect and snRNA/scRNA sequence and analyze GT biopsies relative to snRNAseq reference set.
- Clone dominant TCRs and screen for specificity to dystrophin micro-dystrophin or exon skipped dystrophin peptide antigens.
Statement of Benefit to California:
Duchenne Muscular Dystrophy leads to significant disability and premature death due to progressive muscle weakness, imposing significant physical and financial consequences on patients and their families. Approximately, 1500 California families are affected by DMD. There is a large unmet need as there are no curative treatments. Studies proposed may lead to novel targets for drug discovery, therapeutic strategies for targeting satellite muscle stem cells, and better gene therapies for DMD.